Deconvoluting the Vascular Adhesome

去卷积血管粘附体

• 批准号: 10548841
• 负责人: Mark HOWARD Ginsberg
• 金额: $ 78.7万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10548841
• 关键词: Accounting; Adhesions; Behavior; Binding Sites; Biochemical; Blood; Blood Platelets; Blood Vessels; Cell Adhesion; Cell Line; Cells; Complex; Cytoplasm; Data; Endothelial Cells; Family; Future; Health; Heterogeneity; Image; Integrins; Leukocytes; Ligation; Mesenchymal; Methods; Molecular; Myocardial Infarction; Output; Process; Proteins; Regulation; Resources; Rod; Signal Transduction; Site; Stroke; Structure; Talin; Testing; Vinculin; Visualization; cell assembly; cell motility; density; dynamical evolution; experimental study; member; novel strategies; physical state; therapeutic target

Summary Integrin-based adhesions are central to the functions of blood and vascular cells. The heterogeneity of adhesions and their dynamic evolution has complicated efforts to study their fine composition, assembly, and disassembly. Our studies have shown that a transition between two adhesion archetypes is controlled by a simple binary molecular switch of vinculin competition with and displacement of RIAM or lamellipodin(Lpd), members of the MRL family, from binding sites on talin's rod domain. Furthermore, we developed methods to image the complex of MRL proteins with integrins and talin (MIT complex) and showed that it formed the tips of “sticky fingers,” cellular protrusions that sense the density of matrix proteins and physical state of the substrate during mesenchymal cell migration. mesenchymal cell migration. We hypothesize that the MIT complex represents one among many distinct modules that contribute to the overall structure and function of integrin- based adhesions. This suggests the paradigm that the integrin adhesome can be analyzed as a dynamic assembly of these modules, which form prior to entry into the adhesions. This concept has enabled us to propose a new approach to studying adhesions by developing methods to purify each module formed prior to integrin ligation. To test this paradigm, we propose to purify 4 such modules and to characterize each for a) the presence of talin-activated integrins b) characterize its protein composition. c) establish its biochemical topology and regulation d) visualize it in living cells and e) evaluate its functions The proposed experiments will generate foundational data for the adhesion field in three ways: a) the integrin adhesome will be re- interpreted as a compendium of modular components; each with its own specific composition. b) The functional studies will identify new regulators of blood and vascular cell adhesion and signaling that may be exploitable as therapeutic targets c) A complete definition of the protein composition of each module will serve as a public resource for the analysis of integrin-based adhesions and will be hypothesis-generating for future studies to understand the regulation of assembly of each module and its functional outputs.

概括 基于整合素的粘附对于血液和血管细胞的功能至关重要。 粘连的异质性及其动态演化使得研究其精细化的工作变得复杂起来。 我们的研究表明，两者之间存在过渡。 粘附原型是由纽蛋白竞争的简单二元分子开关控制的 以及 MRL 家族成员 RIAM 或板脂蛋白 (Lpd) 从结合位点的置换 此外，我们开发了对 MRL 蛋白复合物进行成像的方法。 与整合素和talin（MIT复合物）结合，并表明它形成了“粘性手指”的尖端，细胞 突起在过程中感知基质蛋白的密度和基质的物理状态 我们研究了 MIT 的间充质细胞迁移。 复杂代表了对整体做出贡献的许多不同模块之一 基于整合素的粘附的结构和功能这表明了这一范式。 整合素粘附体可以作为这些模块的动态组装进行分析，这些模块形成于 这个概念使我们能够提出一种新的研究方法。 通过开发纯化整合素连接之前形成的每个模块的方法来测试粘附。 在这个范式中，我们建议纯化 4 个这样的模块并表征每个模块的 a) 存在 建立talin激活的整合素 b) 表征其蛋白质组成 c) 其生物化学。 拓扑和调节 d) 在活细胞中可视化它并 e) 评估其功能 实验将以三种方式生成粘附场的基础数据：a）整合素 粘附体将被重新解释为模块化组件的概要；每个组件都有自己的组件； b) 功能研究将确定新的血液和血管调节剂。 可用作治疗靶点的细胞粘附和信号转导 c) 完整的定义 每个模块的蛋白质组成将作为公共资源用于分析 基于整合素的粘连，将为未来的研究提出假设，以了解 每个模块的组装及其功能输出的调节。